Impact of VKORC1, CYP2C9, and CYP4F2 Polymorphisms on Optimal Warfarin Dose: Does Ethnicity Matters?

BACKGROUND: Conventional anticoagulation with warfarin remains the cornerstone strategy for numerous preventive strategies. It is established that Asian patients require lower warfarin doses than Caucasians potentially attributing to the genetic polymorphism (GP) differences. AREAS OF UNCERTAINTY: The impact of GP on optimal warfarin dose (OWD) in Koreans is unclear when compared with other ethnicities. It is also not well established whether GP linked to OWD in Korean patients to the similar extend as in Chinese, Japanese, and Caucasians. DATA SOURCES: Single-center prospective observational study in Koreans, matched with historic cohorts of other ethnicities. THERAPEUTIC ADVANCES: Clinical characteristics, concomitant medications, OWD, international normalized ratio, and VKORC1, CYP2C9, and CYP4F2 GPs were assessed in consecutive Korean patients. The OWD was defined when patient's international normalized ratio was within target range for at least 3 consecutive times separated by 1 week. We included 133 (mean age 62.6 ± 12.1 years, 49% males) warfarin-treated patients of Korean descend. The mean OWD was 3.30 ± 1.34 (range: 1-9) mg/d. Homozygous wild-type patients required lower OWD (3.1 ± 1.1 mg/d vs. 4.7 ± 1.8 mg/d, P < 0.001) for VKORC1 and higher OWD for both CYP2C9 (3.4 ± 1.3 mg/d vs. 2.3 ± 1.1 mg/d, P = 0.002) and CYP4F2 (3.0 ± 1.2 mg/d vs. 3.4 ± 1.3 mg/d vs. 4.0 ± 1.7 mg/d, P = 0.033) than those carrying heterozygote genes. CONCLUSIONS: Korean patients exhibit different VKORC1, CYP2C9, and CYP4F2 profiles impacting lower OWD in Eastern Asians than required in Caucasians. Universal international OWD guidelines may consider patient ethnicity as a confounder; however, this hypothesis needs further clarification.

An LCI-like protein APC2 protects ginseng root from Fusarium solani infection.

AIMS: We aimed to purify an antimicrobial protein from Bacillus amyloliquefaciens FS6 culture supernatant, verify its antimicrobial activity against Fusarium solani and evaluate its biocontrol potential for ginseng root rot. METHODS AND RESULTS: The antimicrobial protein was purified from FS6 culture supernatant using ammonium sulphate precipitation, anion exchange and gel chromatography. Based on mass spectrometry results, the purified protein was identified as an antimicrobial protein of the LCI family and was designated APC2 . The APC2 recombinant protein expressed in Escherichia coli (BL21) significantly inhibited F. solani and decreased the infection and spread of F. solani in ginseng root. An overexpressing APC2 strain FS6-APC2 was constructed and shown to have enhanced antimicrobial activity compared to the wild-type strain FS6. CONCLUSIONS: The APC2 protein shows strong antimicrobial activity against F. solani, reduces the incidence and severity of ginseng root rot caused by F. solani and exhibits a great biocontrol potential. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports the inhibitory activity of APC2 protein (LCI family) against F. solani and its protective efficacy on ginseng root rot. These findings provide a scientific basis for future research on the biocontrol mechanism, as well as the development and application of FS6.

[The 486th case: chronic diarrhea and orthostatic hypotension].

A 57-year-old man was admitted to hospital with diarrhea for 10 months and dizziness for 4 months. The patient had 1-2 liters watery stool per day, without pyogenic blood or abnormality in gastroenteroscopy examination. The level of hemoglobin and albumin was generally normal, and fasting test was positive. At the same time, he was accompanied with hyperalgesia of lower limbs and orthostatic hypotension. After the discussion of multiple disciplinary teams, the patient was diagnosed with amyloidosis by sural nerve biopsy, myocardial MRI, and the assays of urine immunoelectrophoresis and serum free light chain. Light chain amyloidosis was confirmed after excluded the diagnosis of familial amyloidosis. The patient was improved after courses of chemotherapy with melphalan and dexamethasone.

[Effect of splenectomy on the risk of hepatocellular carcinoma development among patients with liver cirrhosis and portal hypertension: a multi-institutional cohort study].

Objective: To identify whether splenectomy for treatment of hypersplenism has any impact on development of hepatocellular carcinoma(HCC) among patients with liver cirrhosis and hepatitis. Methods: Patients who underwent splenectomy for hypersplenism secondary to liver cirrhosis and portal hypertension between January 2008 and December 2012 were included from seven hospitals in China, whereas patients receiving medication treatments for liver cirrhosis and portal hypertension (non-splenectomy) at the same time period among the seven hospitals were included as control groups. In the splenectomy group, all the patients received open or laparoscopic splenectomy with or without pericardial devascularization. In contrast, patients in the control group were treated conservatively for liver cirrhosis and portal hypertension with medicines (non-splenectomy) with no invasive treatments, such as transjugular intrahepatic portosystemic shunt, splenectomy or liver transplantation before HCC development. All the patients were routinely screened for HCC development with abdominal ultrasound, liver function and alpha-fetoprotein every 3 to 6 months. To minimize the selection bias, propensity score matching (PSM) was used to match the baseline data of patients among splenectomy versus non-splenectomy groups. The Kaplan-Meier method was used to calculate the overall survival and cumulative incidence of HCC development, and the Log-rank test was used to compare the survival or disease rates between the two groups. Univariate and Cox proportional hazard regression models were used to analyze the potential risk factors associated with development of HCC. Results: A total of 871 patients with liver cirrhosis and hypertension were included synchronously from 7 tertiary hospitals. Among them, 407 patients had a history of splenectomy for hypersplenism (splenectomy group), whereas 464 patients who received medical treatment but not splenectomy (non-splenectomy group). After PSM,233 pairs of patients were matched in adjusted cohorts. The cumulative incidence of HCC diagnosis at 1,3,5 and 7 years were 1%,6%,7% and 15% in the splenectomy group, which was significantly lower than 1%,6%,15% and 23% in the non-splenectomy group (HR=0.53,95%CI:0.31 to 0.91,P=0.028). On multivariable analysis, splenectomy was independently associated with decreased risk of HCC development (HR=0.55,95%CI:0.32 to 0.95,P=0.031). The cumulative survival rates of all the patients at 1,3,5,and 7 years were 100%,97%,91%,86% in the splenectomy group,which was similar with that of 100%,97%,92%,84% in the non-splenectomy group (P=0.899). In total,49 patients (12.0%) among splenectomy group and 75 patients (16.2%) in non-splenectomy group developed HCC during the study period, respectively. Compared to patients in non-splenectomy group, patients who developed HCC after splenectomy were unlikely to receive curative resection for HCC (12.2% vs. 33.3%,χ²=7.029, P=0.008). Conclusion: Splenectomy for treatment of hypersplenism may decrease the risk of HCC development among patients with liver cirrhosis and portal hypertension.

[Clinical analysis of 350 cases of pneumoconiosis complicated with spontaneous pneumothorax].

Objective: To explore the clinical features of pneumoconiosis complicated with spontaneous pneumothorax to improve the diagnosis and treatment of this disease. Methods: Analyze the clinical characteristics and treatment of 350 cases of pneumoconiosis complicated with spontaneous pneumothorax in Hunan Prevention and Treatment Institute for Occupational Diseases from May 2016 to May 2018. Results: In 350 patients, 22 cases are pneumoconiosis stage I, accounting for 6.3%, 26 cases are pneumoconiosis stage Ⅱ, accounting for 7.4%, 302 cases were pneumoconiosis stage Ⅲ, accounting for 86.3%.168 cases were recurrent pneumothorax, the recurrence rate was as high as 48%.There were 232 cases occurred in winter and spring, accounting for 66.3%. Chronic obstructive pulmonary disease and pulmonary infection were 54.9% and 47.4%, respectively. 233 patients were treated with basic therapy such as high flow oxygen therapy, with an effective rate of 93.1%. 114 cases were treated with thoracic closed drainage, with an effective rate of 86%. Conclusion: Spontaneous pneumothorax is a common complication of pneumoconiosis with high recurrence rate. According to the different conditions to give different treatments in a timely manner can achieve better results.

[The association of pre-pregnancy body mass and weight gain during pregnancy with macrosomia: a cohort study].

Objective: To examine the association of pre-pregnancy body mass and weight gain during pregnancy with macrosomia. Methods: From January 2015 to December 2015, a total of 20 477 pregnant women were recruited by probabilistic proportional scale sampling with simple randomization in Sichuan, Yunnan and Guizhou Provinces. Basic information of pregnant women, weight gain during pregnancy and weight of newborn were collected. A multiple logistic regression model was used to assess the association between the pre-pregnancy body mass and gestational weight gain indicators with macrosomia. Results: 20 321 mother-infant were included in the final analysis. 20 321 pregnant women were (30.09±4.10) years old and delivered at (39.20±1.29) weeks, among which 12 341 (60.73%) cases were cesarean delivery. The birth weight of 20 321 infants were (3 292.26±431.67) grams, and 970 (4.77%) were macrosomia. The multiple logistic regression model showed that after adjusting for the age of women, compared to the normal weight group in the pre-pregnancy, the overweight and obesity group elevated the risk of macrosomia, with OR (95%CI) about 1.99 (95%CI: 1.69-2.35) and 4.05 (95%CI: 3.05-5.39), respectively. After adjusting for the age, the pre-pregnancy BMI, delivery weeks, delivery mode and infant's gender, compared to the weight-gain appropriate group, higher weight gain rate in the mid-pregnancy and excessive total gestational weight gain elevated the risk of macrosomia, with OR (95%CI) about 1.99 (95%CI: 1.66-2.39) and 1.80 (95%CI: 1.55-2.08), respectively. Conclusion: The overweight before pregnancy, obesity before pregnancy, the rate of weight gain in the second trimester and the high total weight gain during pregnancy could increase the risk of macrosomia.

Hyponatremia in neuromyelitis optica spectrum disorders: Literature review.

Hyponatremia is a potentially serious electrolyte abnormality observed in neuromyelitis optica spectrum disorders (NMOSDs), and its most common cause is syndrome of inappropriate antidiuretic hormone secretion (SIADH). Another potential cause of hyponatremia is cerebral salt-wasting syndrome (CSWS), although CSWS has not previously been reported in NMOSDs. Accurate and early differentiation between SIADH and CSWS is difficult. However, the two conditions have important implications for the selection of therapy. Here, we describe two patients with aquaporin-4 antibody (AQP4-Ab)-positive NMOSDs who developed hyponatremia as a result of CSWS and SIADH, respectively. Additionally, we review all previously reported studies of hyponatremia in patients with NMOSDs and propose several potential pathophysiological mechanisms of hyponatremia. In conclusion, NMOSDs accompanied by hyponatremia are not actually rare, but have previously been given little attention. Furthermore, SIADH should not be the only consideration, before the exclusion of rare but significant CSWS.

135G/C polymorphism in the RAD51 gene and acute myeloid leukemia risk: a meta-analysis.

Numerous studies have evaluated the association between the 135G/C polymorphism in the RAD51 gene and risk of acute myeloid leukemia (AML), but the results have been inconsistent. The aim of this study is to precisely examine the association between the 135G/C polymorphism in the RAD51 gene and AML risk through a meta-analysis. PubMed, Google Scholar, and Web of Science databases were systematically searched to identify relevant studies from their inception to June 2015. Pooled odds ratios (OR) with 95% confidence intervals (95%CI) were calculated using fixed- or random-effect models. A total of 6 case-control studies containing 1432 patients and 2750 controls were used in this meta-analysis, and our results showed no association between the 135G/C polymorphism in the RAD51 gene and AML risk (CC vs GG: OR = 1.67, 95%CI = 0.93-3.02; GC vs GG: OR = 1.24, 95%CI = 0.80-1.92; the dominant model: OR = 1.26, 95%CI = 0.83-1.91; the recessive model: OR = 1.63, 95%CI = 0.90-2.95). No publication bias was found in this study. In summary, the present meta-analysis suggests that the 135G/C polymorphism in the RAD51 gene may not be associated with AML risk. However, further studies with larger cohorts are needed to confirm this conclusion.

Development and characterization of microsatellite loci in Brasenia schreberi (Cabombaceae) based on the next-generation sequencing.

To understand genetic variability of the endangered aquatic herb Brasenia schreberi (Cabombaceae), we describe 31 microsatellite markers obtained using next-generation sequencing. A total of 24 individuals from the population of Jackson Lake, USA, were genotyped for each marker. Twenty-eight markers were polymorphic. The number of alleles per locus ranged from 1 to 9; the observed and expected heterozygosities ranged from 0 to 1 and from 0 to 0.751, respectively. These markers should be useful tools for genetic variation and conservation studies of B. schreberi.

Development of microsatellite markers in the tetraploid fern Ceratopteris thalictroides (Parkeriaceae) using RAD tag sequencing.

To understand the genetic variability of the tetraploid fern Ceratopteris thalictroides (Parkeriaceae), we described 30 polymorphic microsatellite markers obtained using the restriction site-associated DNA (RAD) tag sequencing technique. A total of 26 individuals were genotyped for each marker. The number of alleles per locus ranged from 4 to 10, and the expected heterozygosity and the Shannon-Wiener index ranged from 0.264 to 0.852 and 0.676 to 2.032, respectively. Because these 30 microsatellite markers exhibit high degrees of genetic variation, they will be useful tools for studying the adaptive genetic variation and sustainable conservation of C. thalictroides.

Development of microsatellite markers in Hagenia abyssinica (Bruce) J.F. Gmel, an endangered tropical tree of eastern Africa, using next-generation sequencing.

Hagenia abyssinica (Bruce) J.F. Gmel is an endangered tree species endemic to the high mountains of tropical Africa. We used Illumina paired-end technology to sequence its nuclear genome, aiming at creating the first genomic data library and developing the first set of genomic microsatellites. Seventeen microsatellite markers were validated in 24 individuals. The average number of alleles per locus was 7.6, while the observed and expected heterozygosities ranged from 0.000 to 0.958 and from 0.354 to 0.883, respectively. These polymorphic markers will be used as tools for further molecular studies to facilitate formulation of appropriate conservation strategies for this species.

Development of expressed sequence tag-based microsatellite markers for the critically endangered Isoëtes sinensis (Isoetaceae) based on transcriptome analysis.

Isoëtes sinensis is a critically endangered quillwort. To facilitate studies on the conservation genetics of this species, we developed expressed sequence tag-simple sequence repeat (EST-SSR) markers. A total of 50,063 unigenes were predicted by transcriptome sequencing, 5294 (10.6%) of which significantly matched 3011 Gene Ontology annotations and 2363 were assigned to Kyoto Encyclopedia of Genes and Genomes metabolic pathways. Most of these (2297) were involved in metabolism. A total of 1982 SSR motifs were identified, with trinucleotides being the dominant repeat motif, and 1438 (72.6%) SSR primers were designed. Eighteen randomly selected primer pairs were used to genotype 24 I. sinensis accessions, which confirmed the suitability of these novel markers for molecular studies of I. sinensis. The heterozygosity index value ranged between 0.0799 and 0.9106, while the Shannon-Wiener diversity index value ranged between 0.1732 and 2.5589. The EST-SSRs reported in this study are linked to genic sequences, and are therefore ideal for investigating the evolutionary history of I. sinensis. These markers, together with the large EST dataset generated in this study, will greatly facilitate conservation genetic studies of I. sinensis.

Development of EST-SSR markers in the relict tree Davidia involucrata (Davidiaceae) using transcriptome sequencing.

Davidia involucrata, reputed to be a "living fossil" in the plant kingdom, is a relict tree endemic to China. Extant natural populations are diminishing due to anthropogenic disturbance. In order to understand its ability to survive in a range of climatic conditions and to design conservation strategies for this endangered species, we developed genic simple sequence repeats (SSRs) from mRNA transcripts. In total, 142,950 contigs were assembled. Of these, 30,411 genic SSR loci were discovered and 12,208 primer pairs were designed. Dinucleotides were the most common (77.31%) followed by trinucleotides (16.44%). Thirteen randomly selected primers were synthesized and validated using 24 individuals of D. involucrata. The markers displayed high polymorphism with the number of alleles per locus ranging from 3 to 12 and the observed and expected heterozygosities ranging from 0.083 to 1.0 and 0.102 to 0.69, respectively. This large expressed sequence tag dataset and the novel SSR markers will be key tools in comparative studies that may reveal the adaptive evolution, population structure, and resolve the genetic diversity in this endangered species.

Evaluation of the taxonomic status of water dropwort (Oenanthe, Apiaceae) accessions from East Asia based on nuclear rDNA internal transcribed spacer sequences.

Oenanthe L. is a taxonomically complex genus, several species of which have long been used as vegetables and traditional medicines in East Asia. In order to clarify the taxonomic status of Oenanthe accessions and provide baseline data for the sustainable use of its genetic resources, we examined sequence variations in the internal transcribed spacer (ITS) region of Oenanthe accessions collected from a wide geographical area in China and its neighboring countries. For comparison, ITS sequences in GenBank for almost all currently reported species of Oenanthe were also included in our analyses. Both phylogenetic tree construction methods (Bayesian inference and maximum likelihood) revealed that the accessions tended to cluster into two groups, which were closely related to O. mildbraedii and O. sarmentosa. However, these two species have never been recorded in China or its neighboring countries. Therefore, it seems probable that in our sampled locations, Oenanthe accessions have been given an incorrect name, such as O. javanica. Future studies should carefully check the morphological characteristics of other Oenanthe species and sequence their ITS regions in order to clarify the taxonomic status of the genus.

Common genetic variants on 1p13.2 associate with risk of autism.

Autism is a highly heritable neurodevelopmental disorder, and known genetic variants, mostly rare, account only for a small proportion of cases. Here we report a genome-wide association study on autism using two Chinese cohorts as gene discovery (n=2150) and three data sets of European ancestry populations for replication analysis of top association signals. Meta-analysis identified three single-nucleotide polymorphisms, rs936938 (P=4.49 × 10(-8)), non-synonymous rs6537835 (P=3.26 × 10(-8)) and rs1877455 (P=8.70 × 10(-8)), and related haplotypes, AMPD1-NRAS-CSDE1, TRIM33 and TRIM33-BCAS2, associated with autism; all were mapped to a previously reported linkage region (1p13.2) with autism. These genetic associations were further supported by a cis-acting regulatory effect on the gene expressions of CSDE1, NRAS and TRIM33 and by differential expression of CSDE1 and TRIM33 in the human prefrontal cortex of post-mortem brains between subjects with and those without autism. Our study suggests TRIM33 and NRAS-CSDE1 as candidate genes for autism, and may provide a novel insight into the etiology of autism.

Cloning and sequence analysis of the LOC339524 gene in Sprague-Dawley rats.

We cloned the LOC339524 gene in Sprague-Dawley (SD) rats and analyzed the structure and function of the protein encoded by it. Based on the known human LOC339524 gene sequences, the full-length coding sequence of the LOC339524 gene in SD rats was cloned and amplified by the polymerase chain reaction using the complementary DNA of SD rats as a template. Bioinformatics analysis showed that the length of the cloned LOC339524 gene (GenBank accession No. KM224520) was 831 bp and it encoded a deduced protein of 276 amino acids. Sequence analysis revealed that the coded protein was identical to that produced in humans and its functional domain was located in the 138-236 amino acid fragments, a proline-rich region. Our results suggest that the encoded protein may be a significant regulator of the inflammatory response and may provide sufficient information to justify an in-depth investigation of the role of the LOC339524 gene.

Predicting transient particle transport in enclosed environments with the combined computational fluid dynamics and Markov chain method.

To quickly obtain information about airborne infectious disease transmission in enclosed environments is critical in reducing the infection risk to the occupants. This study developed a combined computational fluid dynamics (CFD) and Markov chain method for quickly predicting transient particle transport in enclosed environments. The method first calculated a transition probability matrix using CFD simulations. Next, the Markov chain technique was applied to calculate the transient particle concentration distributions. This investigation used three cases, particle transport in an isothermal clean room, an office with an underfloor air distribution system, and the first-class cabin of an MD-82 airliner, to validate the combined CFD and Markov chain method. The general trends of the particle concentrations vs. time predicted by the Markov chain method agreed with the CFD simulations for these cases. The proposed Markov chain method can provide faster-than-real-time information about particle transport in enclosed environments. Furthermore, for a fixed airflow field, when the source location is changed, the Markov chain method can be used to avoid recalculation of the particle transport equation and thus reduce computing costs.

Effect of mitochondrial ATP-sensitive potassium channel opening on the translocation of protein kinase C epsilon in adult rat ventricular myocytes.

This study aimed to investigate the effects of mitochondrial ATP-sensitive potassium (MitoKATP) channel opening on the translocation of protein kinase C epsilon (PKCε). In addition, we aimed to determine the relationship between PKCε translocation and the production of reactive oxygen species (ROS). PKCε protein expression in cultured adult rat ventricular myocytes was investigated by immunofluorescence and Western blotting. Diazoxide (DZ), a selective MitoKATP channel activator, caused a significant translocation to myofibrillar-like structures in cultured adult rat ventricular myocytes. N-2-Mercaptopropionylglycine, a free radical scavenger, could partially inhibit the translocation of PKCε induced by DZ. By contrast, chelerythrine, a selective PKC inhibitor, could completely block the translocation of PKCε induced by DZ. The opening of MitoKATP channels might activate and cause PKCε to translocate into myofibrillar-like structures. PKCε activation occurred downstream of the MitoKATP channel, possibly as a result of ROS production that occurred after the MitoKATP channels opened.